Method of combatting morphine addiction and toxification

ABSTRACT

Deoxycytidine has been found to be a morphine antagonist. Morphine (heroin) addiction and toxification can be alleviated and combatted by administering deoxycytidine to the addicted or toxified subject. Satisfactory results are obtained by injecting deoxycytidine intra-peritoneally in the amount in the range 151200 mg/kg of body weight, depending on the subject.

United States Patent [191 Penn [ METHOD OF COMBATTING MORPHINE ADDICTIONAND TOXIFICATION [75] Inventor: Nathar W. Penn, Wyckoff, NJ.

[73] Assignee: Addiction Research and Treatment Corporation, Brooklyn,NY.

[22] Filed: Oct. 10, 1973 [21] Appl. No.: 405,025

[52] US. Cl. 424/180 [51] Int. Cl. A0ln 9/00, AOln 9/28 [58] Field ofSearch 424/180; 260/209 1 Mar. 25, 1975 Primary Eraminer-Elbert RobertsAttorney, Agent, or Firm-Cooper, Dunham, Clark, Griffin & Moran [57]ABSTRACT Deoxycytidine has been found to be a morphine z1ntugonist.Morphine (heroin) addiction and toxil'icution can be alleviated andcombutted by administering deoxycytidine to the addicted or toxifiedsubject. Sutisfactory results are obtained by injecting deoxycytidineintra-peritoneally in the amount in the range l5l200 mg/kg of bodyWeight, depending on the sub- Ject.

6 Claims, N0 Drawings METHOD OF COMBATTING MORPHINE ADDICTION ANDTOXIFICATION This invention relates to morphine addiction. Moreparticularly, this invention relates to the treatment of morphine(heroin) addicts and to morphine detoxification.

Morphine (heroin) addiction is a serious social problem. In addition,morphine addiction is an ever present danger in medical practice when amorphine containing drug is administered to a patient to relieve pain.In view of the above, it is highly desirable for social and medicalreasons to have available compositions and techniques employing the sameuseful in overcoming or combatting or alleviating morphine addiction,morphine toxification and combatting withdrawal symptoms experienced bymorphine addicts and others upon withdrawal from morphine.

Accordingly, it is an object of this invention to provide a method ofcombatting morphine or heroin addiction. It is another object of thisinvention to provide a method of alleviating the withdrawal symptomsexperienced by a morphine or heroin addict upon withdrawal from themorphine or heroin.

In at least one embodiment of the practice of this invention at leastone of the foregoing objects will be achieved.

It has been discovered that deoxycytidine is a morphine antagonist.Deoxycytidine is employed as a morphine antagonist by administeringdeoxycytidine, preferably in a physiologically acceptable carrier, suchas in a non-pyrogenic aqueous solution or suspension, to the subjectsuffering from morphine or heroin addiction or toxification or sufferingfrom morphine or heroin withdrawal symptoms. Deoxycytidine isadministered intraperitoneally, but other routes of administration maybe employed, such as orally. Usually, an amount of deoxycytidine in therange about -1200 mg/kg of body weight of the subject being treatedyields satisfactory results, although more or less may be employed. Inthe usual practice, an amount in the range 75-500 mg/kg, e.g. 100 mg/kg,would be employed.

In the practices of this invention it is preferred to employdeoxycytidine derived from mammalian DNA. Deoxycytidine is obtained frommammalian DNA by the breakdown of the DNA with pancreatic DNAase. Theresulting breakdown products are then subjected to another enzymictreatment involving alkaline phosphatase to produce deoxycytidine andother deoxynucleosides. The resulting products are thenchromatographically fractionated to yield deoxycytidine. In all of thesesteps the pH is held in the range from about 3.5 to about 9.

In these tests carried out to demonstrate the value and utility of thepractices of this invention animals were employed. Animals, like man,are subject to morphine addiction and toxification and also experiencewithdrawal symptoms. The use of animal models in studies of morphineaddiction is recognized and accepted and generally success in theseanimal studies point to and indicates a substantial equivalent successor effect in man.

In tests carried out in animals (mice) it was found that deoxycytidineantagonized the Straub effect and hyperactivity within about 2 or 3minutes. In contrast the compound cytidine exhibited no activity.Deoxycytidine was shown to be effective in antagonizing the analgesiceffect of morphine in rats as gauged by the flinch-jump test. Thenaloxone-jump assay indicated the effectiveness and activity ofdeoxycytidine in detoxifying morphine treated mice.

In tests carried out to demonstrate the effectiveness of deoxycytidineas a morphine antagonist pairs of rats weighing about 300 grams eachwere injected intraperitoneally with morphine in an amount of about 20mg/kg of animal body weight. The animals response to graded shock wastested one half hour later, and the voltages required to elicit flinchand jump reactions were noted. Deoxycytidine in the amount mg/kg of bodyweight was then injected into one of each of the pairs of the animalstested and the pairs were then tested again after one hour. Thedifferences in shock strength required to elicit the flinch reactionwere then determined. There was some decline in shock intensity whichcaused flinching normally occurring both in the morphine-treatedcontrols and in the untreated animals. There was, however, a significantdifference observed between deoxycytidine-treated morphinized animalsand those which received morphine alone.

The naloxone jump assay is a measure of the severity of the withdrawalsyndrome or degree of physical dependence on morphine. A significantdifference (p 0.05) in the number of naloxone-induced jumps shown bydeoxycytidine-treated morphine tolerantdependent mice compared with theuntreated animals was observed.

The substitution of cytidine for deoxycytidine abolishes this mitigationof withdrawal symptoms. The effect is pharmacologically specific todeoxycytidine, which differs from cytidine only in one hydroxyl group ofthe molecule. Deoxycytidine in one massive dose thus acts as aneffective detoxicant in morphine tolerant-dependent mice. Other testsshows that deoxycytidine effected a moderate stimulation of thedepressed respiratory rate in a morphinized dog. Although there is aspecies variation in the response of morphinetreated animals todeoxycytidine, the tests shown deoxycytidine to be a morphineantagonist. For example, deoxycytidine when administered to amorphinized rabbit did not produce as great an increase in respiratoryrate as in the dog.

The observed variability in response might be due to differences totissue levels of endogenous factors involved or associated withdeoxycytidine in its antagonist action. It is theorized that when theamount of these unknown factors are high or present, deoxycytidine actseffectively as an antagonist. At low levels, however, it may be thatthese low levels result in a show of less activity on the part ofdeoxycytidine as an antagonist.

The use of deoxycytidine as a morphine antagonist is especiallyattractive since it is non-toxic, non-addictive and a compound which canreadily be prepared. These properties of deoxycytidine provide majoradvantages for the use of this compound as a morphine (heroin)antagonist for the treatment of drug addicts and morphine (heroin)detoxification. The fact that deoxycytidine is non-addictive andcellularly endogenous makes the subject or addict disposed to theadministration of deoxycytidine in a treatment regime.

It is believed that the drug addict would have a more positive responsewith respect to treatment by deoxycytidine which stresses, in effect,the restoration of normal cell processes since deoxycytidine treatmentwould be comparable or similar to vitamin or hormonal treatment.

In the test referred to hereinabove the quantitative determination ofthe antagonism of morphine analgesic action was performed with apparatuswhich delivers an electrical shock to rats [Evans, W. 0.,Psychopharmacologia 2:319 (1961)]. Strength of the shock was varied insteps by the operator; the equipment automatically delivers voltage to agrid housed in a box with a glass inset at the top. The shock occurredevery 30 seconds for a one second duration. It was scrambled, that is,different bars in the grid are electrified each time so that the ratdoes not learn which are ground and which have applied voltage. Theminimal strength of shock at which the animal flinches and at which itjumps was determined by successive up and down stepwise variations ofshock strength.

In carrying out these tests morphine sulfate was injectedintraperitoneally (i?) into rats at the dosage mg/kg. The flinch levelwas determined /2 hour later. Experience has shown that maximal effectof morphine occurs at this time and the literature reports that thistime corresponds to the maximal concentration of morphine in the brainfor this species under these conditions (Mule, S. .l., in NarcoticDrugs: Biochemical Pharmacology, ed. D. H. Clouet, p. 106, Plenum Press,NY. 1971). As soon as the determination was completed, deoxycytidine,120 mg/kg in saline solution, or saline, or cytidine, 128 mg/kg insaline solution was injected IP. The latter two solutions were controls.The flinch shock threshhold was determined again after 1 hour. Thecontrols normally showed a decrease in the shock strength required toelicit a flinch reaction due to decline in the morphine effect. Thechanges were compared with the changes in threshold for thedeoxycytidine-treated animals using the unpaired Student t test. Thisstandard statistical test gave p 0.0l, indicating a highly significantdifference between the test group and controls, whether the latter aresaline or cytidine.

Another experiment was carried out to show that deoxycytidine does not,of itself, affect the flinch threshhold in normal animals. In this case,the changes in flinch threshhold are small, since morphine was notinjected and there was no wearing-off effect. It was found thatdeoxycytidine had no action on the flinch threshhold in thenon-narcotized rat. The observed changes were, therefore, due to itsantagonism of morphine, not to a generalized stimulation. These data aresummarized in accompanying Table 1.

TABLE 1 EFFECT OF DEOXYC'YTlDlNE TCBAEATMENT ON CHANGE OF Rats injectedIP with morphine sulfate, 20 mg/kg or with saline, llinch threshholddetermined Vzh later. Animals then injected IP with deoxycytidine, I20mg/kg saline. or cylidine 128 mg/kg. threshhold tested again after 1Mean :se, p .01 for difference between morphine-deoxycytidine group andcontrols; no significant difference in threshhold changes between groupsgiven saline followed by deoxycytidine or saline.

In many species morphine severely depresses respiratory rate. This is amajor cause of death due to overdosage in humans. Deoxycytidine willincrease the rate of respiration in those morphine-treated animals whichshow this markedly lowered respiration in response to morphine.Deoxycytidine stimulates the respiration in both the morphine-treateddog and rabbit. The effect is more evident and prolonged in the former.This may, in part, be due to a species difference in response, but isprobably also related to the dosages of morphine required to produce thelowered respiratory rate. In the dog, 4.9 mg/kg was injected lP, whereasmg/kg was given the rabbits. It is known that the functions of cellularcomponents other than deoxycytidine or deoxycytidine-containingcompounds are affected by morphine (Dodge, P. W. and Takemori, A. E.,Biochem. Pharmacol. 21:287, 1972). The apparent diminished efficiency ofdeoxycytidine in the rabbit may, therefore, actually result frommorphine effects on other constituents of the cell due to theconsiderable dosage employed. In both species, deoxycytidine atapproximately three times the weight of morphine was used forantagonism. In the dog, the morphinized respiratory rate wasapproximately doubled for the period of morphine action. In the rabbit,the increase was to about 50-75% of normal, varying over about 2 hoursor from Ax- A1 of the duration of the morphine effect in this species.

An animal which can be shown to require morphine as a result ofprolonged administration is termed tolerant-dependent to avoidanthropomorphic coloration. Standard procedures have been devised toinduce this condition in various species. The mouse is most convenientand a schedule of increasing morphine dosage has been reported in theliterature with a quantitative test for the degree of morphinedependence (Marshall, I. and Weinstock M., Nature 2341223, 1971 Micewere injected IP daily for 8 days, starting at 50 mg/kg and increasingthe dosage by this amount each day. On the eighth day, naloxone, 50mg/kg, was given IP 4 hours after the morphine injection. Naloxone is amorphine antagonist which induces jumping in a morphine-dependent (oraddicted) animal. The number of jumps in a standard period of time is ameasure of the degree of dependence (addiction). If a separateantagonist were used prior to naloxone, its effectiveness iscounter-acting the dependent (addictive) condition would be determinedquantitatively by a reduction in the number of jumps. The overallprocedure corresponds to the process of withdrawal in the human, so thata decreased number ofjumps indicates an amelioration of the withdrawalsymptoms. The foregoing is standard pharmacological interpretation. Theresults of such an assay are given in accompanying Table 2.Deoxycytidine given at a dosage of 3 times the weight of the finalmorphine injection significantly reduces the number of jumps by thetreated group. It, therefore, acts to mitigate the withdrawal symptomsin this standard assay.

TABLE 2 NALOXONE INDUCED .IUMPS IN DEOXYCYTIDINE- TREATED AND CONTROLMORPHlNE-DEPENDENT MICE* Male Swiss albino mice were injected IP withmorphine sulfate. 50 mg/kg. dosage increased daily by this amount forseven days to the level of 400 mg/kg.

" Mean 1: s.e. naloxone. 50 mg/kg. given lP alternately to individualanimals of test and control groups 4 h after final morphine injection.Deoxycytitline. 1.2 gm/kg. cytidine 1.28 gm/kg or saline given torespective groups Ah before naloxone.

In other tests mice were injected 1? with morphine sulfate at the levelof 55 mg/kg. In about 5-10 minutes the tail of the mouse became raisedupright or arched forward over the back. This is known as the Straubeffect. At the same time the mice began to run aimlessly about the cageshowing an extreme hyperactivity. This assay differs from the precedingone in that animals are not tolerant-dependent. It is termed an assay ofthe morphine acute effect. At this point deoxycytidine at the level of165 mg/kg was injected. The mice began to become calm after 5-10 minutesand the upright tails returned to a normal position. In the next 2hours. there were fitful spurts of morphine symptoms but, on theaverage. a qualitative difference in appearance and behavior wasevident.

Other test data are set forth in accompanying Table 3.

TABLE 3 MILLIAMPERAGE FOR THRESHHOLD FLICK* Initial Final Test. Morphinedeoxycytidine .882 .457

Control, Morphine H 0 Untreated NUMBER OF JUMPS IN EIGHT-MINUTES PERIODAFTER NALOXONE INJECTION Test Control Application ofthe t test gives p0.()2 for the difference in response between the two groups.

As will be apparent to those skilled in the art in the light of theforegoing disclosure, many modifications, alterations and substitutionsare possible in the practice of this invention without departing fromthe spirit or scope thereof.

I claim:

1. A method for treatment of morphine addiction, or morphinetoxification and morphine withdrawal symptoms which comprisesadministering to the subject suffering the same an effective amount of acomposition consisting essentially of deoxycytidine.

2. A method in accordance with claim 1 wherein the deoxycytidine isadministered intraperitoneally.

3. A method in accordance with claim I wherein said deoxycytidine isadministered intravenously.

4. A method in accordance with claim 1 wherein the deoxycytidine isadministered in an amount in the range 15-1200 mg/kg per body weight perthe subject.

5. A method in accordance with claim 4 wherein said deoxycytidine isadministered in an amount in the range mg/kg subject body weight.

6. A method in accordance with claim 1 wherein said deoxycytidine isadministered in a non-pyrogenic aqueous physiologically acceptablecarrier therefor.

UNITED STATES PATENT OFFICE i CERTIFICATE OF CORRECTION P atent No.3,873,698 Dated March 25, 1975 Iriventofls) Nathar Ww Penn It iscertified that error appears in the above-identified patent and thatsaid Letters Patent are hereby corrected as shown below:

The assignee should correctly read:

Nathar W. Penn Claim 1, line 1, "or" should be omitted.

Column 2, line 46, "to" should correctly read-- in Signed and Sealedthis twenty-second Day Of July 1975 [SEAL] v Atresr:

RUTH c. MASON c. MARSHALL DANN Arresting Officer Commissioner of Patentsand Trademark:

1. A METHOD FOR TREATMENT OF MORPHINE ADDICTION, OR MORPHINETOXIFICATION AND MORPHINE WITHDRAWAL SYMPTOMS WHICH COMPRISESADMINISTERING TO THE SUBJECT SUFFERING THE SAME AN EFFECTIVE AMOUNT OF ACOMPOSITION CONSISTING ESSENTIALLY OF DEOXYCLYTIDINE.
 2. A method inaccordance with claim 1 wherein the deoxycytidine is administeredintraperitoneally.
 3. A method in accordance with claim 1 wherein saiddeoxycytidine is administered intravenously.
 4. A method in accordancewith claim 1 wherein the deoxycytidine is administered in an amount inthe range 15-1200 mg/kg per body weight per the subject.
 5. A method inaccordance with claim 4 wherein said deoxycytidine is administered in anamount in the range 100 mg/kg subject body weight.
 6. A method inaccordance with claim 1 wherein said deoxycytidine is administered in anon-pyrogenic aqueous physiologically acceptable carrier therefor.